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J. Med. Microbiol. -- Vol. 50 (2001), 795-804
© 2001 Society for General Microbiology
ISSN 0022-2615

BACTERIAL PATHOGENICITY

Identification of two Mycobacterium avium subspecies paratuberculosis gene products differentially recognised by sera from rabbits immunised with live mycobacteria but not heat-killed mycobacteria

JOHN P. BANNANTINE* and JUDITH R. STABEL

National Animal Disease Center, ARS-USDA, Ames, IA 50010, USA

Corresponding author: Dr J.P. Bannantine (e-mail: jbannant{at}nadc.ars.usda.gov).

Received 6 Nov. 2000; revised version accepted 5 May 2001.

Abstract

The investigation of environmentally regulated proteins has led to a better understanding of host–pathogen interactions and identified novel vaccine candidate antigens for several bacterial pathogens. In an effort to identify such proteins in Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis), a genomic expression library was differentially screened with sera from rabbits that had been immunised with live M. paratuberculosis ({alpha}-live) as well as sera from rabbits immunised with heat-killed M. paratuberculosis ({alpha}-killed). These experiments identified seven recombinant plaques that were uniquely recognised by the {alpha}-live sera. Sequence data showed that five of these clones overlapped with each other and contained a common open-reading frame encoding a 25-kDa protein, termed Csp1. The 25-kDa antigen shows weak similarity to a secreted Corynebacterium glutamicum protein. The remaining two clones overlapped with each other and contained two partial open-reading frames, both encoding proteins with strong homology to polyketide synthase from various species of mycobacteria. Antisera were produced against a peptide of the polyketide synthase gene product designated Pks7. Csp1-specific antibodies were affinity purified from the {alpha}-live sera. These purified antibodies demonstrated that Csp1 was present within infected macrophages. Collectively, these data identify novel M. paratuberculosis antigens that may be important in pathogenesis.




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